中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (12): 2994-3004.doi: 10.12307/2026.661

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

关节康片干预软骨稳态保护膝骨关节炎小鼠关节软骨

李宜金1，黎嘉澔2，张海涛3，黄艺伟4，陈锦伦5，曾意荣5，冯文俊5   

  1. 1广州中医药大学深圳中西医结合临床医学院，广东省深圳市  518100；2广州番禺区中医院关节骨科，广东省广州市  511400；3上海中医药大学附属龙华医院，上海市  200032；4广州中医药大学附属中山中医院，广东省中山市  528400；5广州中医药大学第一附属医院，广东省广州市  510405
  • 收稿日期:2025-03-13 接受日期:2025-08-04 出版日期:2026-04-28 发布日期:2025-09-28
  • 通讯作者: 冯文俊，博士，副教授，广州中医药大学第一附属医院，广东省广州市 510405 共同通讯作者：曾意荣，博士，教授，广州中医药大学第一附属医院，广东省广州市 510405
  • 作者简介:李宜金，男，1995年生，河南省南阳市人，汉族，广州中医药大学在读博士，主要从事膝骨关节炎及骨质疏松的机制及中医药防治研究。
  • 基金资助:
    国家自然科学基金青年基金项目(82104882)，项目负责人：冯文俊；国家自然科学基金面上项目(82374484)，项目负责人：曾意荣

GJK Tablets intervene in cartilage homeostasis to protect articular cartilage of mice with knee osteoarthritis

Li Yijin1, Li Jiahao2, Zhang Haitao3, Huang Yiwei4, Chen Jinlun5, Zeng Yirong5, Feng Wenjun5   

  1. Li Yijin1, Li Jiahao2, Zhang Haitao3, Huang Yiwei4, Chen Jinlun5, Zeng Yirong5, Feng Wenjun5
    1Shenzhen Clinical College of Integrated Chinese and Western Medicine, Guangzhou University of Chinese Medicine, Shenzhen 518100, Guangdong Province, China; 2Department of Joint Orthopedics, Panyu District Hospital of Traditional Chinese Medicine, Guangzhou 511400, Guangdong Province, China; 3Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China; 4Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine, Zhongshan 528400, Guangdong Province, China; 5The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2025-03-13 Accepted:2025-08-04 Online:2026-04-28 Published:2025-09-28
  • Contact: Zeng Yirong, MD, Professor, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Li Yijin, MD candidate, Shenzhen Clinical College of Integrated Chinese and Western Medicine, Guangzhou University of Chinese Medicine, Shenzhen 518100, Guangdong Province, China Corresponding author: Feng Wenjun, MD, Associate professor, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    The National Natural Science Foundation of China (Youth Fund), No. 82104882 (to FWJ); The National Natural Science Foundation of China (General Program), No. 82374484 (to ZYR)

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摘要：


文题释义：
软骨稳态：软骨通过正常合成代谢与分解代谢维持软骨稳定性，衰老或创伤等原因诱导的软骨细胞退变、凋亡及软骨细胞外基质过度降解，可造成软骨稳态失衡，进展为软骨损伤。
SRY-Box转录因子9(Sox9)：是Sox转录因子家族的成员，该家族的特征是高迁移率族 (HMG)-box DNA结合域。 Sox9在整个胚胎的间充质凝聚早期表达，是软骨发育和骨骼形成过程中必不可少的软骨促进因子。

背景：既往研究发现，关节康片能够改善膝骨关节炎患者关节疼痛及功能，但具体机制尚未明确。 
目的：探讨关节康片对膝骨关节炎模型小鼠软骨保护的作用机制。
方法：网络药理学方法获取关节康片和软骨稳态的共同靶点，构建蛋白质相互作用网络，通过GO分析及KEGG富集分析筛选关键信号通路。动物实验将小鼠随机分为对照组、模型组、关节康低、中、高剂量组、盐酸氨基葡萄糖胶囊组(阳性药物)，后5组小鼠采用改良Hulth法构建小鼠膝骨关节炎模型，对照组小鼠手术仅暴露相关组织，不切除韧带及半月板。造模成功后关节康低、中、高剂量组分别予187.5，375，750 mg/kg关节康片混悬液灌胃，盐酸氨基葡萄糖胶囊组予盐酸氨基葡萄糖混悬液187.5 mg/kg灌胃，模型组、对照组予等量生理盐水灌胃，隔日1次给药，共8周。灌胃结束后，通过苏木精-伊红和番红O-固绿染色观察小鼠膝关节软骨病理形态，并进行软骨OARSI评分，免疫组化染色检测小鼠软骨组织中Sox9、Ⅱ型胶原蛋白、基质金属蛋白酶13的蛋白表达。
结果与结论：①共获得关节康片与软骨稳态交集靶点140个，Sox9为靶点之一；②GO分析和KEGG富集分析显示主要涉及的通路有癌症相关通路、缺氧诱导因子1、MAPK、PI3K/Akt、核因子κB、转化生长因子β等信号通路；③苏木精-伊红染色及番红O-固绿染色结果显示，与对照组相比，模型组软骨轮廓形变，缺损分层，软骨细胞数量明显减少，潮线模糊，OARSI评分高；与模型组相比，关节康各剂量组软骨表面完整，软骨厚度及软骨细胞数量增加，潮线清晰，OARSI评分均明显降低；④免疫组化结果提示，与对照组相比，模型组Sox9、Ⅱ型胶原蛋白表达明显降低，基质金属蛋白酶13表达明显升高(P < 0.01)；与模型组相比，关节康各剂量组基质金属蛋白酶13表达明显减少(P < 0.01)；关节康片中、高剂量组的Sox9、Ⅱ型胶原蛋白阳性表达均明显提高(P < 0.01)，且从低剂量到高剂量组阳性表达呈增加趋势(P < 0.01)；⑤结果表明，关节康片可通过调控Sox9干预软骨稳态，在膝骨关节炎中发挥软骨保护作用，缺氧诱导因子1、MAPK、PI3K/Akt、核因子κB、转化生长因子β等信号通路可能是关节康片干预软骨稳态的主要通路。

关键词: 关节康片, 膝骨关节炎, 软骨稳态, 关节软骨, Sox9

Abstract: BACKGROUND: Previous studies have found that GJK Tablets can improve joint pain and function in patients with knee osteoarthritis, but the specific mechanism remains unclear.
OBJECTIVE: To explore the mechanism underlying the chondroprotective effect of GJK Tablets in a mouse model of knee osteoarthritis.
METHODS: Network pharmacology was used to obtain the common targets of GJK Tablets and cartilage homeostasis, and a protein-protein interaction network was constructed. Key signaling pathways were screened through gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis. Experimental mice were randomly divided into a control group, a model group, low-, medium-, and high-dose GJK Tablet groups, and a positive drug group (glucosamine hydrochloride capsules). In the latter five groups, the modified Hulth method was used to establish the mouse model of knee osteoarthritis. In the control group, only the relevant tissues of mice were exposed but the ligaments and menisci were not removed. After modeling, the low, medium-, and high-dose groups were intragastrically administered with 187.5, 375, and 750 mg/kg GJK Tablet suspension, respectively. The positive drug group was intragastrically administered with 187.5 mg/kg glucosamine hydrochloride suspension. The model group and the control group were intragastrically administered with an equal volume of normal saline. The administration was carried out every other day for 8 weeks. After the intragastric administration, the pathological morphology of the knee joint cartilage of the mice was observed by hematoxylin-eosin and safranin O-fast green staining, and the Osteoarthritis Research Society International (OARSI) score for the cartilage was evaluated. The protein expressions of SRY-Box transcription factor 9 (Sox9), type II collagen, and matrix metalloproteinase 13 in the mouse cartilage tissue were detected by immunohistochemical staining.
RESULTS AND CONCLUSION: (1) A total of 140 intersection targets of GJK Tablets and cartilage homeostasis were obtained, and Sox9 was one of the targets. (2) Gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that the main signaling pathways involved were cancer-related pathways, hypoxia-inducible factor 1, MAPK, PI3K/Akt, nuclear factor κB, transforming growth factor β and other signaling pathways. (3) The results of hematoxylin-eosin staining and safranin O-fast green staining showed that compared with the control group, the cartilage contour in the model group was deformed, the defect was stratified, the number of chondrocytes was significantly reduced, the tidemark was blurred, and the OARSI score was higher. Compared with the model group, the cartilage surface in the low-, medium-, and high-dose GJK Tablet groups was more intact, the cartilage thickness and the number of chondrocytes were increased, the tidemark was clearer, and the OARSI scores were significantly decreased. (4) The immunohistochemical results indicated that compared with the control group, the protein expressions of Sox9 and type II collagen in the model group were significantly decreased, and the expression of matrix metalloproteinase 13 was significantly increased (P < 0.01). Compared with the model group, the expression of matrix metalloproteinase 13 in the low-, medium-, and high-dose GJK Tablet groups was significantly decreased (P < 0.01). The positive expressions of Sox9 and type II collagen in the medium- and high-dose GJK Tablet groups were significantly increased (P < 0.01), and the positive expressions showed an increasing trend from the low-dose group to the high-dose group (P < 0.01). To conclude, GJK Tablets can intervene in cartilage homeostasis by regulating Sox9, playing a chondroprotective role in knee osteoarthritis. Signaling pathways such as hypoxia-inducible factor 1, MAPK, PI3K/Akt, nuclear factor κB, and transforming growth factor β may be the main pathways through which GJK Tablets intervene in cartilage homeostasis. 

Key words: GJK Tablets, knee osteoarthritis, cartilage homeostasis, articular cartilage, Sox9

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